Kawamura Akinori, Yamada Kouji, Fujimori Ken'ichi, Higashinakagawa Toru
Department of Biology, School of Education, Waseda University, 1-6-1 Nishi-Waseda, Shinjuku, Tokyo 169-8050, Japan.
Biochem Biophys Res Commun. 2002 Feb 22;291(2):245-54. doi: 10.1006/bbrc.2002.6447.
Here we describe isolation and characterization of two zebrafish cDNAs, designated ph2alpha and ph2beta, which were identified as structural homologs of the Drosophila polyhomeotic, mouse Mph2, and human HPH2 genes, collectively termed the Polycomb group. The alpha and beta transcripts shared a 1.9-kb sequence at their 3'-termini. Alpha had an additional 1.6-kb sequence extending toward its 5'-terminus. Only a short 0.1-kb segment was unique to beta. Sequencing of a genomic clone corresponding to the two cDNAs indicated that the mRNAs were transcribed from a single gene locus by alternative promoters. Northern blots revealed expression of alpha transcripts during the segmentation period, while beta expression occurred at all developmental stages examined. Whole-mount in situ hybridizations with an alpha-specific probe and a probe recognizing both transcripts revealed distinct spatio-temporal expression patterns along developing somites. Alpha transcripts were detected initially at the 7-8 somite stage; beta transcripts appeared in the first somites. As segmentation proceeded, alpha and beta expression shifted position toward the tailbud in parallel with the formation of each somite. Within individual somites, the signal corresponding to alpha was strongest at the posterior border and weakest in the anterior region. Conversely, that corresponding to beta was strongest at the anterior border and weakest in the posterior region. The data support the idea that Ph2alpha and Ph2beta are involved in spatio-temporal generation of somites as well as in specification of antero-posterior regional differences within individual somites.
在此,我们描述了两个斑马鱼cDNA(分别命名为ph2alpha和ph2beta)的分离与特性分析,它们被鉴定为果蝇多同源异型基因、小鼠Mph2基因和人类HPH2基因的结构同源物,这些基因统称为多梳蛋白家族。α和β转录本在其3'末端共享一段1.9 kb的序列。α在其5'末端还延伸有一段额外的1.6 kb序列。β独有的只有一段短的0.1 kb片段。对与这两个cDNA对应的基因组克隆进行测序表明,这些mRNA是由单个基因位点通过可变启动子转录而来。Northern杂交显示α转录本在体节形成期表达,而β在所有检测的发育阶段均有表达。用α特异性探针和能识别两种转录本的探针进行的全胚胎原位杂交揭示了沿发育中的体节的不同时空表达模式。α转录本最初在7 - 8体节期被检测到;β转录本出现在最初的体节中。随着体节形成过程的进行,α和β的表达位置与每个体节的形成同步向尾芽移动。在单个体节内,与α对应的信号在后部边界最强,在前部区域最弱。相反,与β对应的信号在前部边界最强,在后部区域最弱。这些数据支持了Ph2alpha和Ph2beta参与体节的时空生成以及单个体节内前后区域差异的特化这一观点。